Salmon Louse Lepeophtheirus Salmonis Research Articles

Insights from Hi-C data regarding the Pacific salmon louse (Lepeophtheirus salmonis) sex-chromosomes.

Salmon lice, Lepeophtheirus salmonis (family Caligidae), are ectoparasites that have negatively impacted the salmon aquaculture industry and vulnerable wild salmon populations. Researchers have studied salmon lice to better understand their biology to develop effective control strategies. In this study, we updated the chromosome-level reference genome assembly of the Pacific subspecies of L. salmonis using Hi-C data. The previous version placed contigs/scaffolds using an Atlantic salmon louse genetic map. By utilizing Hi-C data from Pacific salmon lice, we were able to properly assign locations to contigs/scaffolds previously unplaced or misplaced. This resulted in a more accurate genome assembly and a more comprehensive characterization of the sex-chromosome unique to females (W). We found evidence that the same ZW-ZZ mechanism is common in both Atlantic and Pacific subspecies of salmon lice using PCR assays. The W-chromosome was approximately 800 kbp in size, which is ∼30 times smaller than the Z-chromosome (24 Mbp). The W-chromosome contained 61 annotated genes, including 32 protein-coding genes, 27 long non-coding RNA (lncRNA) genes, and 2 pseudogenes. Among these 61 genes, 39 genes shared homology to genes found on other chromosomes, while 20 were unique to the W-chromosome. Two genes of interest on the W-chromosome, prohibitin-2 and kinase suppressor of ras-2, were previously identified as potential sex-linked markers in the salmon louse. However, we prioritized the 20 unique genes on the W-chromosome as sex-determining candidates. This information furthers our understanding of the biology of this ectoparasite and may help in the development of more effective management strategies.

Effect of diel light cycles on vertical migration patterns of Lepeophtheirus salmonis (Krøyer, 1837) copepodids measured in an in situ mesocosm

AbstractThe ectoparasitic salmon louse (Lepeophtheirus salmonis Krøyer, 1837) persists as a major hindrance for continued growth of the Norwegian salmon industry. Shielding technologies or management strategies are being used to reduce the likelihood of contact in the net pens between the salmon and the sea lice’s infectious copepodid stage. Knowledge about the vertical distribution and diel migration patterns of the planktonic sea lice stages are inevitably a premise for such water-depth-dependent avoidance technologies to be efficient; however, not many studies have investigated diel vertical migration patterns of sea lice in situ or in relevant scales. In this work, we aimed to monitor the diel migration of L. salmonis copepodids in the vertical plane without the influence of hydrographical forces, using sufficient water column depth of 11 m, natural thermoclines, and natural exogenous light conditions. We followed a cohort of copepodids over the course of 4 days and measured their vertical distribution in the water column twice per hour in a custom made mesocosm, using automatic particle detection and machine learning to analyse position and migration patterns. The L. salmonis copepodid population displayed reverse diel migration, and we found a statistically significant interaction between depth and time. During the day, copepodids were found in the upper 1 m of the water column, before the majority of the population descended to 1–2 m depth during night-time. A small fraction of the population remained in the upper part of the water column also during night-time. A significantly higher fraction of the copepodid population resided in the upper metre during daylight hours compared to other depths. These results could be utilised as input in sea lice dispersal models which are important management tools in today’s sea lice prevention strategies and regulatory frameworks.

Comparison of DNA metabarcoding and morphological diet analysis of lumpfish (Cyclopterus lumpus) as methods of estimating sea lice cleaning efficacy inside commercial Atlantic salmon (Salmo salar) sea cages

Lumpfish (Cyclopterus lumpus) are often used as cleaner fish inside salmonid sea cages for biological control of ectoparasitic sea lice. We compared traditional morphological diet assessments with DNA metabarcoding as methods of estimating lumpfish cleaning efficacy as a function of its size, and of its ingestion of alternative prey. Traditional morphological analysis was performed on 122 ethanol-preserved stomachs. The stomachs were removed in the field - following eDNA protocols – from lumpfish collected from industrial Atlantic salmon sea cages in southern Newfoundland. Morphological stomach analysis gave semi-quantitative estimates of prey counts and/or biomass at the order or family level; it showed that shorter lumpfish ate more gammarid amphipods whereas longer lumpfish ate more formulated feed pellets designed for salmon. No sea lice were confidentially morphologically identified in the lumpfish stomachs. In contrast, COI metabarcoding was able to confidently identify the presence of the salmon louse (Lepeophtheirus salmonis) in 61 of 82 successfully barcoded lumpfish stomachs. Metabarcoding showed that diurnally-migrating krill and gammarid amphipods were present in more stomachs in 2018 whereas formulated feed pellets – identified from COI sequences classified as domestic poultry and non-indigenous fish species - were present in more stomachs in 2020. We argue that, when weighed by the total number of reads, the counts per stomach of the number of Global Estimated Sequence Variants (ESV) classified as a particular prey species could be used to estimate the single-species indices of abundance needed for lumpfish breeding values. The number of ESVs classified as the salmon louse could be used to estimate an index of lumpfish cleaning efficacy. As proof-of-concept, we show that 83% of the variance in the number of Norwegian krill heads observed per stomach was explained by two weighed covariates: 1) Norwegian krill ESV counts and 2) lumpfish stomach contents weight.

Salmon louse labial gland enzymes: implications for host settlement and immune modulation.

Salmon louse (Lepeophtheirus salmonis) is a skin- and blood-feeding ectoparasite, infesting salmonids. While feeding, labial gland proteins from the salmon louse may be deposited on the Atlantic salmon (Salmo salar) skin. Previously characterized labial gland proteins are involved in anti-coagulation and may contribute to inhibiting Atlantic salmon from mounting a sufficient immune response against the ectoparasite. As labial gland proteins seem to be important in the host-parasite interaction, we have, therefore, identified and characterized ten enzymes localized to the labial gland. They are a large group of astacins named L. salmonis labial gland astacin 1-8 (LsLGA 1-8), one serine protease named L. salmonis labial gland serine protease 1 (LsLGSP1), and one apyrase named L. salmonis labial gland apyrase 1 (LsLGAp1). Protein domain predictions showed that LsLGA proteins all have N-terminal ShK domains, which may bind to potassium channels targeting the astacins to its substrate. LsLGA1 and -4 are, in addition, expressed in another gland type, whose secrete also meets the host-parasite interface. This suggests that LsLGA proteins may have an anti-microbial function and may prevent secondary infections in the wounds. LsLGAp1 is predicted to hydrolyze ATP or AMP and is, thereby, suggested to have an immune dampening function. In a knockdown study targeting LsLGSP1, a significant increase in IL-8 and MMP13 at the skin infestation site was seen under LsLGSP1 knockdown salmon louse compared to the control, suggesting that LsLGSP1 may have an anti-inflammatory effect. Moreover, most of the identified labial gland proteins are expressed in mature copepodids prior to host settlement, are not regulated by starvation, and are expressed at similar or higher levels in lice infesting the salmon louse-resistant pink salmon (Oncorhynchus gorbuscha). This study, thereby, emphasizes the importance of labial gland proteins for host settlement and their immune dampening function. This work can further contribute to anti-salmon louse treatment such as vaccine development, functional feed, or gene-edited salmon louse-resistant Atlantic salmon.

Histopathological assessment of Atlantic salmon exposed to calcium oxide particles: a controlled clinical study

Particulate calcium oxide (CaO) has been successfully used for the control of sea urchin and starfish populations. Recent studies have proven its efficacy in killing planktonic copepods, making it a promising and cheap option for salmon louse control in the Atlantic salmon (Salmo salar) farming industry. Additionally, preliminary laboratory tests demonstrated that exposure to 0.2 g/L–0.6 g/L of fine CaO particles induced significant mortality of salmon louse (Lepeophtheirus salmonis) in the planktonic stages. The present study investigated the effects of water treatment with fine CaO particles [0.2 g/L or 127.4 g/m2; 0.1 mm–0.3 mm], conducted twice a week for three consecutive weeks at 5°C and 12°C on Atlantic salmon post-smolt in a flow-through system. The study compared mortality rates and histopathology of skin, eyes, gills, and intestines between treated and untreated control salmon. The results indicated that CaO exposure did not induce fish mortality or histopathological damages in the skin, eyes, or intestines. Although there were no significant effects of CaO exposure on gill inflammation or hyperplasia, the exposure did increase the occurrence of gill vascular injuries and necrosis in small portions of the tissue (< 10% of the respiratory gill tissue in most of the samples) by 60% and 35%, respectively. The effect was not modulated by temperature or time. The results indicate that the CaO treatment protocol used in this study is not lethal to Atlantic salmon and is safe for most analyzed tissues. Nonetheless, the induction of vascular damage and necrosis in the gill, albeit in small tissue portions, emphasizes the need for further research. It is crucial to develop a protocol that avoids such lesions, ensuring the secure application of CaO as an antiparasitic treatment in salmon aquaculture.

Exploring the impact of thermal delousing on gill health and microbiome dynamics in farmed Atlantic Salmon

The economic cost of salmon louse (Lepeophtheirus salmonis and Caligus rogercresseyi) infestations in the Atlantic salmon (Salmo salar) industry has been estimated to be around $900 million annually. This high cost has driven a concerted effort to develop, examine, understand, and implement various methods for louse control. Husbandry interventions utilising warm water exposure have been highly successful in complementing traditional chemotherapeutants, especially as the efficiency of the latter has reduced in recent years. In this study, we sought to examine the impact of thermal delousing on gill health in two commercial sites with different historical husbandry and treatment interventions prior to and post-thermal treatment. Methods to characterise gill health and the response to thermal treatment included a detailed examination of the gill for microparasites using both histology and qPCR and targeted immune gene expression analysis, most notably antigen-presenting cells (mhc ii), proinflammatory cytokines (il-1β and tnf-α) and inhibitory cytokines (tgf-β and il-10). Furthermore, we examined the bacterial communities present on the gill surface using 16S rRNA amplicon sequencing. Data obtained from these trials indicated a minimal impact on gill microparasite prevalence in response to the thermal treatment. The expression of immune markers exhibited a significant decrease across both sites after treatment. Intriguingly, marked differences in the gill bacterial communities in response to treatment between the two sites were clearly observed. This divergence could be attributed to the notable differences in husbandry history and health status of the fish at the two sites prior to the thermal treatment. Our data suggest that microbiome diversity is an informative indicator of fish gill health and could be used to define appropriate interventions when treating sea lice.

Evolutionary predictions for a parasite metapopulation: Modelling salmon louse resistance to pest controls in aquaculture.

Pests often evolve resistance to pest controls used in agriculture and aquaculture. The rate of pest adaptation is influenced by the type of control, the selective pressure it imposes, and the gene flow between farms. By understanding how these factors influence evolution at the metapopulation level, pest management strategies that prevent resistance from evolving can be developed. We developed a model for the metapopulation and evolutionary dynamics of the salmon louse (Lepeophtheirus salmonis), which is a major parasite affecting salmon aquaculture. Different management scenarios were simulated across a network of salmon farms covering half of Norway, and their effects on louse epidemiology and evolution were investigated. We compared louse controls that differed in how they were deployed through time (discrete vs. continuous), how they impacted the louse life cycle, and in their overall efficacy. We adjusted the strength of selection imposed by treatments, the dominance effect of the resistant allele, and the geographic location at which resistance originated. Continuously acting strategies (e.g., louse-resistant salmon) were generally more effective than discrete strategies at controlling lice, especially when they increased louse mortality during early developmental stages. However, effective strategies also risked imposing frequent and/or strong selection on lice, thus driving rapid adaptation. Resistant alleles were more likely to be lost through genetic drift when they were recessive, had a low-fitness advantage, or originated in low-farm-density areas. The north-flowing current along the Norwegian coastline dispersed resistant genes from south to north, and limited gene flow in the opposite direction. We demonstrate how evolutionary models can produce quantitative predictions over large spatial and temporal scales and for a range of pest control scenarios. Quantitative outputs can be translated into practical management decisions applied at a regional level to minimise the risk of resistance developing.

The digestion time for salmon louse (Lepeophtheirus salmonis) in lumpfish (Cyclopterus lumpus)

Atlantic salmon aquaculture employs lumpfish as a control method to combat ectoparasites, given their unique cleaning behaviour. There are multiple studies which estimate the average number of salmon lice in the stomach contents of dissected lumpfish. However, these numbers cannot be used to assess the cleaning efficacy of lumpfish (e.g., the average number of lice consumed daily per lumpfish) without knowing the digestion time of lice in lumpfish. The aim of the study was to provide quantitative estimates of the degradation of salmon lice, through a blinded clinical study over a duration of seven days. Individually tagged lumpfish (45.8 g, SD ± 10.28) were randomly arranged in triplicate tanks (n = 28 per tank) and acclimatised for three days. Subsequently, lumpfish were fed using oral gavage dosing with counts of lice (0–6), feed pellets (0–6) or a combination of both. Lice used were recently captured and stored at −80 °C to prevent parasite transmission at the study location and photographed before and after digestion to estimate degradation. Samplings ranged from 6 h intervals during the first two days, to 24 h and eventually 48 h for the last two days. Analysis of salmon lice revealed an expected digestion time of 29 h while the median digestion time was estimated to 15 h at 9 °C. Pellets dissolved quickly and had no impact on the lice digestion time. The findings of this study can be used to estimate cleaning efficacy of lumpfish from stomach contents of salmon lice.

De novo high-accuracy transcriptomes from long-read sequencing reveals a wide variety of novel splice variants in copepodids and adult female salmon lice (Lepeophtheirus salmonis)

Former transcriptome studies of the ectoparasitic salmon louse (Lepeophtheirus salmonis) are based on short-read sequencing and in silico predictions, with the disadvantage of inadequately describing splice variants and insufficient differentiation between duplicated genes. In the present study, a de novo full-length transcriptome (TSA accession GKKU00000000) was generated using single-molecule long-read RNA-sequencing (PacBio IsoSeq platform) corrected by short reads (Illumina platform) from the same RNA samples. The two samples, cephalothorax of an adult female and her copepodid offspring, were analyzed separately to facilitate comparison and identification of transcripts unique to each life stage. Each transcript has been supported by two or more full-length non-chimeric reads and at least three short reads, ensuring high-sequence accuracy. A total of 31,092 unique high-accuracy full-length transcripts with an open reading frame > 150 bp, originating from 10,034 unique loci of the salmon louse genome, were identified. More than half of the transcripts are life-stage specific, exclusively present in either the copepodid or adult sample. Approximately one-third of the transcripts were full splice matches with predicted protein coding transcripts presented in NCBI, thus validating these. More than half of the transcripts constituted novel isoforms with at least one new splicing site. We conclude that the full-length transcriptomes represent a versatile reference resource of transcripts. Suitable applications include expression studies, SNP mining, and studies on the biological effects of differences in gene (or isoform) expression between copepodids and adult females. The additional functional annotation of 88% of transcripts allows for identification of gene families of particular interest and for exploration of gene networks and enrichment analysis following expression studies.

Aquaculture-driven evolution of the salmon louse mtDNA genome.

Resistance toward the antiparasitic pyrethroid, deltamethrin, is reported in the Atlantic salmon louse (Lepeophtheirus salmonis salmonis), a persistent ectoparasite of farmed and wild salmonids. The resistance mechanism is linked to mitochondrial DNA (mtDNA), where genetic markers for resistance have been identified. Here, we investigated how widespread pyrethroid use in aquaculture may have influenced mtDNA variation in lice, and the dispersion of resistant haplotypes across the North Atlantic, using historical (2000-2002 "pre-resistance") and contemporary (2014-2017 "post-resistance") samples. To study this, we sequenced ATPase 6 and cytochrome b, genotyped two genetic markers for deltamethrin resistance, and genotyped microsatellites as "neutral" controls of potential population bottlenecks. Overall, we observed a modest reduction in mtDNA diversity in the period 2000-2017, but no reduction in microsatellite variation was observed. The reduction in mtDNA variation was especially distinct in two of the contemporary samples, fixed for one and two haplotypes, respectively. By contrast, all historical samples consisted of close to one mtDNA haplotype per individual. No population genetic structure was detected among the historical samples for mtDNA nor microsatellites. By contrast, significant population genetic differentiation was observed for mtDNA among some of the contemporary samples. However, the observed population genetic structure was tightly linked with the pattern of deltamethrin resistance, and we therefore conclude that it primarily reflects the transient mosaic of pyrethroid usage in time and space. Two historically undetected mtDNA haplotypes dominated in the contemporary samples, both of which were linked to deltamethrin resistance, demonstrating primarily two origins of deltamethrin resistance in the North Atlantic. Collectively, these data demonstrate that the widespread use of pyrethroids in commercial aquaculture has substantially altered the patterns of mtDNA diversity in lice across the North Atlantic, and that long-distance dispersion of resistance is rapid due to high level of genetic connectivity that is observed in this species.

Modelling parasite impacts of aquaculture on wild fish: The case of the salmon louse (Lepeophtheirus salmonis) on out-migrating wild Atlantic salmon (Salmo salar) smolt.

For effective wild salmon (Salmo salar) conservation in areas where aquaculture of salmon is practiced it is necessary to identify where the key parasite, the salmon louse (Lepeophtheirus salmonis), will have an impact on these wild salmon. A simple modelling structure is implemented in a sample system in Scotland for assessing interaction between wild salmon and salmon lice from salmon farms. The model is demonstrated for case studies of smolt sizes and migration routes through salmon lice concentration fields derived for average farm loads from 2018 to 2020. Lice modelling describes production and distribution of lice, infection rates on hosts and biological development of lice. The modelling framework allows explicit assessment of the relationships between lice production, lice concentration and impact on hosts as they grow and migrate. Lice distribution in the environment is determined using a kernel model, which summarises mixing in a complex hydrodynamic system. Smolt modelling describes their initial size, growth and migration pathways. This is illustrated for a set of parameter values applied to 10cm, 12.5cm and 15cm salmon smolts. We found that salmon lice impact depends on initial size of host, smaller smolts will be more susceptible, while larger smolts are less impacted by a given number of lice encounters and migrate more rapidly. This modelling framework can be adapted to allow evaluation of threshold concentrations of lice in the water that should not be exceeded to avoid impacts on smolt populations.

The skin mucosal barrier of lumpfish (Cyclopterus lumpus L.) is weakened by exposure to potential aquaculture production-related stressors.

Various cleaner fish species, such as the lumpfish (Cyclopterus lumpus L.), are used in the sea cage production of Atlantic salmon (Salmo salar L.) as a control measure against the ectoparasitic salmon louse (Lepeophtheirus salmonis). Nonetheless, during severe lice infestation, alternative treatments are required to control parasitic burden. The aim of this study was to gain insight into how lumpfish skin responds to different chemicals used to treat parasites. The authors collected skin from lumpfish from both research facilities (tank-reared fish) and commercial production (cage-reared fish) and used operational welfare indicators, in vitro models, histology and transcriptomics to study how the skin responded to two anti-parasitic oxidative chemicals, hydrogen peroxide (H2 O2 ) and peracetic acid. Lumpfish sampled from the farm were classified as clinically healthy or weak based on their morbidity status, and fish from each category were used to gain insight into how the therapeutics affect the skin barrier. Differences between healthy and weakened (moribund) fish, and between treated fish from each of the two groups, were observed. Histological examination showed an overall reduced skin quality in fish characterized as moribund, including different grades of exposed bony plates. In vitro oxidant-treated lumpfish skin had reduced the migration capacity of keratocytes, a weakened epidermal barrier, and altered gene transcription, changes that are known predisposing factors to secondary infections. Skin from non-treated, healthy fish sampled from commercial farms exhibited similar features and attributes to oxidant-exposed tank-reared fish from a research facility, suggesting that apparently healthy cage-held lumpfish exhibited stress responses in the epidermal barrier. The results of the study outline the risks and consequences lumpfish can face if accidentally subjected to potential anti-parasitic oxidant treatments aimed at Atlantic salmon. It also strengthens the evidence behind the requirement that lumpfish should be removed from the cages before being potentially exposed to this type of treatment and outlines the potential risks of differing husbandry practices upon lumpfish health, welfare and resilience.

In sea trout, the physiological response to salmon louse is stronger in female than in males.

The aims of this study were to compare male and female sea trout (Salmo trutta) with respect to their hypo-osmoregulatory ability over a simulated migration to seawater and their clinical and physiological response to salmon louse (Lepeophtheirus salmonis) infection in seawater and over a simulated pre-mature return to freshwater. For this purpose, 2-year-old hatchery-reared male and female brown trout (F1 offspring of wild caught anadromous fish) were infected with salmon lice and measured for changes in plasma ions, glucose, lactate and osmolality and relative heart, liver and gonad sizes during a simulated seawater migration and thereafter a premature return to freshwater after 4weeks in seawater (pre-adult louse). Un-infected trout served as control. Male trout used longer time to develop full hypo-osmoregulatory ability in seawater and showed a stronger response in plasma glucose and lactate following simulated premature return to freshwater, compared to female trout. Response to salmon louse was stronger in female trout, shown by stronger osmotic stress by chalimus (plasma Cl-) and pre-adult louse (plasma osmolality) and elevated relative liver size (hepatosomatic index) by pre-adult louse in female compared to male trout. Moreover, high plasma cortisol in infected female and low plasma cortisol in infected male trout produced a significant treatment-sex interaction on plasma cortisol. Lice infection intensity was initially higher in male (0.18 lice g-1) compared to female trout (0.11 lice g-1) at the chalimus stage, but equal between sexes at the pre-adult stage (male 0.15 and female 0.17 lice g-1). This study showed that female trout were better adapted for changes in water salinity, while male trout were more robust against salmon louse infection. These results suggests that the elevated salmon louse infection pressure generated by salmon farming have strong and unexplored negative effects on wild sea trout populations. Further research on this topic is vital for the conservation of wild sea trout populations.

Environmentally responsive parasite prevention halves salmon louse burden in commercial marine cages

Though a naturally occurring species throughout the Atlantic, parasitic salmon louse (Lepeophtheirus salmonis) population dynamics are increasingly intertwined with salmonid aquaculture. In contrast to reactive louse management, tools and strategies which reduce louse infestation success provide an off-ramp from the 'more farming equals more lice' feedback loop. This experiment tested the efficacy of a dynamic, environmentally responsive louse prevention strategy using common, commercially available tools throughout a full production cycle at commercial scale. By strategically luring salmon away from the halocline where concentrations of infective louse copepodids are highest using feeding and lights, and minimizing surface water flow through the cage with a protective skirt barrier when no halocline was present, both new louse infestations and mobile louse numbers were cut by half compared to control cages. The reduced louse numbers resulted in 25% fewer delousing events and improved fish welfare in dynamic cages, with no differences in gill condition or growth between treatments. With farmed salmon driving the ecology of salmon lice, this dynamic, environmentally responsive prevention strategy offers a way to work with nature, rather than against it, to reduce the parasite burden on both farmed and wild salmonids.

Key role of mitochondrial mutation Leu107Ser (COX1) in deltamethrin resistance in salmon lice (Lepeophtheirus salmonis)

The pyrethroid deltamethrin (DTM) is used to treat Atlantic salmon (Salmo salar) against salmon louse (Lepeophtheirus salmonis) infestations. However, DTM resistance has evolved in L. salmonis and is currently common in the North Atlantic. This study aimed to re-assess the association between DTM resistance and mitochondrial (mtDNA) mutations demonstrated in previous reports. Among 218 L. salmonis collected in Scotland in 2018–2019, 89.4% showed DTM resistance in bioassays, while 93.6% expressed at least one of four mtDNA single nucleotide polymorphisms (SNPs) previously shown to be resistance associated. Genotyping at further 14 SNP loci allowed to define three resistance-associated mtDNA haplotypes, named 2, 3 and 4, occurring in 72.0%, 14.2% and 7.3% of samples, respectively. L. salmonis strains IoA-02 (haplotype 2) and IoA-10 (haplotype 3) both showed high levels (~ 100-fold) of DTM resistance, which was inherited maternally in crossing experiments. MtDNA haplotypes 2 and 3 differed in genotype for 17 of 18 studied SNPs, but shared one mutation that causes an amino acid change (Leu107Ser) in the cytochrome c oxidase subunit 1 (COX1) and was present in all DTM resistant while lacking in all susceptible parasites. We conclude that Leu107Ser (COX1) is a main genetic determinant of DTM resistance in L. salmonis.

Changes in the Skin Proteome and Signs of Allostatic Overload Type 2, Chronic Stress, in Response to Repeated Overcrowding of Lumpfish (Cyclopterus lumpus L.)

Lumpfish (Cyclopterus lumpus L) is a North Atlantic species harvested for its roe and increasingly used as a cleanerfish in Atlantic salmon (Salmo salar L.) farming to remove salmon louse (Lepeophtheirus salmonis). In aquaculture, the health and welfare of fish depends on optimal levels of several biotic and abiotic factors. Crowding, a common abiotic stress factor in aquaculture practice, can affect the welfare and survival of fish. In this study, lumpfish was exposed to crowding stress daily at random timepoints for one month (stress group) or no crowding (control group). Blood and skin were sampled weekly for physiological parameter analysis and proteomics, respectively. Adrenocorticotropic hormone (ACTH) stimulation and dexamethasone (DEX) suppression test were conducted at the sampling timepoints. Gel-based proteomics coupled with liquid chromatography and tandem mass spectrometry (LC-MS/MS) was used to identify protein changes in skin tissues of lumpfish under crowding. The results indicated that the stress group showed signs of allostatic overload type 2 (chronic stress) due to oversensitivity to ACTH, and a reduced negative feedback system with increased baseline levels of cortisol. These chronic changes in the endocrine system promoted changes in secondary and tertiary stress responses as reduced osmoregulatory capacity and stunted growth, after 14 days of stress and onward. Calmodulin, guanine nucleotide binding protein subunit beta 2, glutathione-S-transferase Mu 3, fatty acid binding protein, heat shock cognate 70 kDa protein, keratin, histone H4 and 14-3-3 alpha/beta showed protein spot intensity changes compared with controls in lumpfish skin at one or several time points during the one month period of crowding stress. The differentially expressed proteins are related to several metabolic pathways and are involved in stress and immune responses. Overall, the study shows that lumpfish can suffer from chronic stress with possible dire consequences for the animal welfare.

Small, charged proteins in salmon louse (Lepeophtheirus salmonis) secretions modulate Atlantic salmon (Salmo salar) immune responses and coagulation

Little is known about glandular proteins secreted from the skin- and blood-feeding ectoparasite salmon louse (Lepeophtheirus salmonis). The labial gland has ducts extending into the oral cavity of the lice, and the present study aimed to identify novel genes expressed by this gland type and to investigate their role in modulation of host parameters at the lice feeding site. Five genes associated with labial gland function were identified and named Lepeophteirus salmonis labial gland protein (LsLGP) 1–4 and 1 like (LsLGP1L). All LsLGPs were predicted to be small charged secreted proteins not encoding any known protein domains. Functional studies revealed that LsLGP1 and/or LsLGP1L regulated the expression of other labial gland genes. Immune dampening functions were indicated for LsLGP2 and 3. Whereas LsLGP2 was expressed throughout the parasitic life cycle and found to dampen inflammatory cytokines, LsLGP3 displayed an increased expression in mobile stages and appeared to dampen adaptive immune responses. Expression of LsLGP4 coincided with moulting to the mobile pre-adult I stage where hematophagous feeding is initiated, and synthetic LsLGP4 decreased the clotting time of Atlantic salmon plasma. Results from the present study confirm that the salmon louse secretes immune modulating and anti-coagulative proteins with a potential application in new immune based anti-salmon louse treatments.

An update of the salmon louse (Lepeophtheirus salmonis) reference genome assembly.

Salmon lice have plagued the salmon farming industry and have negatively impacted salmon populations in the wild. In response, researchers have generated high density genetic maps, genome assemblies, transcriptomes, and whole-genome resequencing data to better understand this parasite. In this study, we used long-read sequencing technology to update the previous genome assemblies of Atlantic Ocean salmon lice with a more contiguous assembly and a more comprehensive gene catalog of Pacific Ocean salmon lice. We were also able to further characterize genomic features previously identified from other studies by using published resequenced genomes of 25 Atlantic and 15 Pacific salmon lice. One example was further characterizing the ZW sex chromosomes. For both the Atlantic and Pacific Ocean salmon lice subspecies, we found that the female W-chromosome is only a small fraction of the Z-chromosome and that the vast majority of the W and Z-chromosome do not contain conserved regions (i.e. pseudoautosomal regions). However, conserved orthologous protein sequences can still be identified between the W- and Z-chromosomes.

Seasonal temperature regulates network connectivity of salmon louse

Abstract Chronically high infestation of salmon louse (Lepeophtheirus salmonis) questions the sustainability of the Norwegian Atlantic salmon (Salmo salar) aquaculture industry. The confinement of millions of hosts, within hundreds of farms with overlapping larval dispersal kernels create the structure for extremely persistent parasite meta-populations. However, the processes regulating the temporal variation in cross-contamination of pelagic salmon louse stages among farms (i.e. connectivity), a vital process driving louse population dynamics, are not well described. Here, we employ a data driven biophysical dispersal model that reproduces three-and-a-half years of production histories of 132 salmon farms in western Norway and quantifies the connectivity of infective pelagic lice stages among the farms with the ocean currents. We show that although the complex geography of western Norwegian fjords governs the long-term topology of the connectivity network, there was a strong seasonal component to network fragmentation. The main de-structuring agent was the delayed infectivity of the pelagic lice stages at cooler temperatures increasing dispersal distances, enhanced by occasional large scale wind forcing events. Coordinated fallowing strategies and de-lousing treatments only played a marginal role in network fragmentation, suggesting that novel lice restraining strategies that consider the environmentally sensitive transport distances must be developed to successfully break up the connectivity network.

Larval salmon lice Lepeophtheirus salmonis exhibit behavioral responses to conspecific pre-adult and adult cues.

In the larval stage of the parasitic copepod Lepeophtheirus salmonis, the free-living copepodid must locate and settle on a salmonid host. Chemosensory mechanisms play a role in determining whether a potential host is suitable for attachment, yet the full suite of chemical cues and resulting behavioral mechanisms used are unknown. After maturing, pre-adult female and adult male salmon lice aggregate upon salmonid hosts for reproduction. Copepodid salmon lice have been observed preferentially infesting hosts that harbor conspecific adults. The aim of this study was to investigate the possibility that salmon lice copepodids perceive and respond to cues from pre-adult female, adult-male, and/or gravid female salmon lice. Behavioral bioassays were conducted in vitro with copepodids exposed to water conditioned with 3 stages of conspecific lice (pre-adult female, adult male, or gravid female), and seawater conditioned with Atlantic salmon Salmo salar Linnaeus, 1758. Experiments demonstrated that copepodids exposed to water conditioned with the salmon host, pre-adult female, or adult male salmon lice significantly altered their behavior, whereas salmon lice exposed to water conditioned with gravid females did not. These results are potentially valuable in the development of novel methods for mitigation of L. salmonis in the salmon aquaculture industry.